Aspirin's antithrombotic effect is mediated predominately by inhibition of platelet cyclooxygenase-1, leading to a decline in serum thromboxane A2 concentrations. We performed a placebo-controlled, randomized, double-blind trial to determine whether aspirin could be given at 3-day intervals and still achieve potent serum thromboxane inhibition. One hundred nine healthy men and women with no recent exposure to aspirin and no contraindications to its use participated. Subjects received 325 mg, 81 mg, or 40 mg of plain aspirin every third day, with placebo on other days; 81 mg of aspirin every day; or placebo every day. Serum concentrations of thromboxane B2 (the metabolite of thromboxane A2) were measured at 3-day intervals during a 31-day treatment period, as well as 4, 7, and 14 days after treatment ended. Serum thromboxane B2 concentrations were nearly identical during treatment with 325 mg of aspirin every third day or 81 mg of aspirin per day (86% inhibition [84%, 89%] and 85% inhibition [73%, 96%], respectively). An aspirin dose of 81 mg every third day was nearly as potent (74% inhibition [70%, 79%]), whereas 40 mg of aspirin every third day achieved only 50% inhibition (40%, 60%). Every-third-day low-dose aspirin regimens (325 and 81 mg) deserve comparison with daily low-dose aspirin regimens in controlled clinical trials because the former regimens could prove to have equal efficacy with reduced toxicity.